{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T02:41:17Z","timestamp":1787020877846,"version":"build-2736575974"},"reference-count":74,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2024,3,16]],"date-time":"2024-03-16T00:00:00Z","timestamp":1710547200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2024,3,16]],"date-time":"2024-03-16T00:00:00Z","timestamp":1710547200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100009088","name":"Veterin\u00e4rmedizinische Universit\u00e4t Wien","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100009088","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Vet Res"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Epithelial damage due to gastrointestinal disorders frequently causes severe disease in horses. To study the underlying pathophysiological processes, we aimed to establish equine jejunum and colon enteroids (eqJE, eqCE) mimicking the in vivo epithelium. Therefore, enteroids were cultivated in four different media for differentiation and subsequently characterized histomorphologically, on mRNA and on protein level in comparison to the native epithelium of the same donor horses to identify ideal culture conditions for an in vitro model system. With increasing enterocyte differentiation, the enteroids showed a reduced growth rate as well as a predominantly spherical morphology and less budding compared to enteroids in proliferation medium. Combined or individual withdrawal of stem cell niche pathway components resulted in lower mRNA expression levels of stem cell markers and concomitant differentiation of enterocytes, goblet cells and enteroendocrine cells. For eqCE, withdrawal of Wnt alone was sufficient for the generation of differentiated enterocytes with a close resemblance to the in vivo epithelium. Combined removal of Wnt, R-spondin and Noggin and the addition of DAPT stimulated differentiation of eqJE at a similar level as the in vivo epithelium, particularly with regard to enterocytes. In summary, we successfully defined a medium composition that promotes the formation of eqJE and eqCE consisting of multiple cell types and resembling the in vivo epithelium. Our findings emphasize the importance of adapting culture conditions to the respective species and the intestinal segment. This in vitro model will be used to investigate the pathological mechanisms underlying equine gastrointestinal disorders in future studies.<\/jats:p>","DOI":"10.1186\/s13567-024-01283-0","type":"journal-article","created":{"date-parts":[[2024,3,16]],"date-time":"2024-03-16T06:01:54Z","timestamp":1710568914000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["A matter of differentiation: equine enteroids as a model for the in vivo intestinal epithelium"],"prefix":"10.1186","volume":"55","author":[{"given":"Christina","family":"Windhaber","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna","family":"Heckl","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Georg","family":"Csukovich","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Barbara","family":"Pratscher","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Iwan Anton","family":"Burgener","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nora","family":"Biermann","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2178-2270","authenticated-orcid":false,"given":"Franziska","family":"Dengler","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,3,16]]},"reference":[{"key":"1283_CR1","doi-asserted-by":"publisher","first-page":"448","DOI":"10.1111\/j.2042-3306.1997.tb03157.x","volume":"29","author":"MK Tinker","year":"1997","unstructured":"Tinker MK, White NA, Lessard P, Thatcher CD, Pelzer KD, Davis B, Carmel DK (1997) Prospective study of equine colic incidence and mortality. Equine Vet J 29:448\u2013453","journal-title":"Equine Vet J"},{"key":"1283_CR2","doi-asserted-by":"publisher","first-page":"204","DOI":"10.1016\/j.prevetmed.2011.06.002","volume":"101","author":"JL Ireland","year":"2011","unstructured":"Ireland JL, Clegg PD, McGowan CM, Platt L, Pinchbeck GL (2011) Factors associated with mortality of geriatric horses in the United Kingdom. Prev Vet Med 101:204\u2013218","journal-title":"Prev Vet Med"},{"key":"1283_CR3","doi-asserted-by":"publisher","first-page":"343","DOI":"10.1111\/j.1939-1676.2003.tb02459.x","volume":"17","author":"MA van der Linden","year":"2003","unstructured":"van der Linden MA, Laffont CM, van Sloet Oldruitenborgh-Oosterbaan MM (2003) Prognosis in equine medical and surgical colic. J Vet Intern Med 17:343\u2013348","journal-title":"J Vet Intern Med"},{"key":"1283_CR4","doi-asserted-by":"publisher","first-page":"2760","DOI":"10.3748\/wjg.v22.i9.2760","volume":"22","author":"J Grootjans","year":"2016","unstructured":"Grootjans J, Lenaerts K, Buurman WA, Dejong CHC, Derikx JPM (2016) Life and death at the mucosal-luminal interface: new perspectives on human intestinal ischemia-reperfusion. World J Gastroenterol 22:2760\u20132770","journal-title":"World J Gastroenterol"},{"key":"1283_CR5","doi-asserted-by":"publisher","first-page":"1183","DOI":"10.1016\/j.bbamcr.2017.03.007","volume":"1864","author":"S Lechuga","year":"2017","unstructured":"Lechuga S, Ivanov AI (2017) Disruption of the epithelial barrier during intestinal inflammation: quest for new molecules and mechanisms. Biochim Biophys Acta Mol Cell Res 1864:1183\u20131194","journal-title":"Biochim Biophys Acta Mol Cell Res"},{"key":"1283_CR6","doi-asserted-by":"publisher","first-page":"157","DOI":"10.1146\/annurev-animal-030117-014748","volume":"6","author":"A Blikslager","year":"2018","unstructured":"Blikslager A, Gonzalez L (2018) Equine intestinal mucosal pathobiology. Annu Rev Anim Biosci 6:157\u2013175","journal-title":"Annu Rev Anim Biosci"},{"key":"1283_CR7","doi-asserted-by":"publisher","first-page":"33","DOI":"10.1016\/j.jcmgh.2017.03.007","volume":"4","author":"E Martini","year":"2017","unstructured":"Martini E, Krug SM, Siegmund B, Neurath MF, Becker C (2017) Mend your fences: the epithelial barrier and its relationship with mucosal immunity in inflammatory bowel disease. Cell Mol Gastroenterol Hepatol 4:33\u201346","journal-title":"Cell Mol Gastroenterol Hepatol"},{"key":"1283_CR8","doi-asserted-by":"publisher","first-page":"340","DOI":"10.1007\/s11894-010-0130-3","volume":"12","author":"S Umar","year":"2010","unstructured":"Umar S (2010) Intestinal stem cells. Curr Gastroenterol Rep 12:340\u2013348","journal-title":"Curr Gastroenterol Rep"},{"key":"1283_CR9","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1007\/s10565-005-0085-6","volume":"21","author":"Y Sambuy","year":"2005","unstructured":"Sambuy Y, de Angelis I, Ranaldi G, Scarino ML, Stammati A, Zucco F (2005) The Caco-2 cell line as a model of the intestinal barrier: influence of cell and culture-related factors on Caco-2 cell functional characteristics. Cell Biol Toxicol 21:1\u201326","journal-title":"Cell Biol Toxicol"},{"key":"1283_CR10","doi-asserted-by":"publisher","first-page":"1073","DOI":"10.1177\/2050640617722903","volume":"5","author":"M Noben","year":"2017","unstructured":"Noben M, Vanhove W, Arnauts K, Santo Ramalho A, van Assche G, Vermeire S, Verfaillie C, Ferrante M (2017) Human intestinal epithelium in a dish: current models for research into gastrointestinal pathophysiology. United European Gastroenterol J 5:1073\u20131081","journal-title":"United European Gastroenterol J"},{"key":"1283_CR11","doi-asserted-by":"publisher","first-page":"248","DOI":"10.1083\/jcb.80.2.248","volume":"80","author":"A Quaroni","year":"1979","unstructured":"Quaroni A, Wands J, Trelstad RL, Isselbacher KJ (1979) Epithelioid cell cultures from rat small intestine. Characterization by morphologic and immunologic criteria. J Cell Biol 80:248\u2013265","journal-title":"J Cell Biol"},{"key":"1283_CR12","doi-asserted-by":"publisher","first-page":"278","DOI":"10.1290\/0404035.1","volume":"40","author":"I Bartsch","year":"2004","unstructured":"Bartsch I, Zschaler I, Haseloff M, Steinberg P (2004) Establishment of a long-term culture system for rat colon epithelial cells. In Vitro Cell Dev Biol Anim 40:278\u2013284","journal-title":"In Vitro Cell Dev Biol Anim"},{"key":"1283_CR13","doi-asserted-by":"publisher","first-page":"G705","DOI":"10.1152\/ajpgi.00518.2003","volume":"288","author":"X-H Weng","year":"2005","unstructured":"Weng X-H, Beyenbach KW, Quaroni A (2005) Cultured monolayers of the dog jejunum with the structural and functional properties resembling the normal epithelium. Am J Physiol Gastrointest Liver Physiol 288:G705\u2013G717","journal-title":"Am J Physiol Gastrointest Liver Physiol"},{"key":"1283_CR14","doi-asserted-by":"publisher","first-page":"241","DOI":"10.1146\/annurev.physiol.010908.163145","volume":"71","author":"LG van der Flier","year":"2009","unstructured":"van der Flier LG, Clevers H (2009) Stem cells, self-renewal, and differentiation in the intestinal epithelium. Annu Rev Physiol 71:241\u2013260","journal-title":"Annu Rev Physiol"},{"key":"1283_CR15","doi-asserted-by":"publisher","first-page":"3759","DOI":"10.1074\/jbc.R114.635995","volume":"291","author":"NC Zachos","year":"2016","unstructured":"Zachos NC, Kovbasnjuk O, Foulke-Abel J, In J, Blutt SE, de Jonge HR, Estes MK, Donowitz M (2016) Human enteroids\/colonoids and intestinal organoids functionally recapitulate normal intestinal physiology and pathophysiology. J Biol Chem 291:3759\u20133766","journal-title":"J Biol Chem"},{"key":"1283_CR16","doi-asserted-by":"publisher","first-page":"262","DOI":"10.1038\/nature07935","volume":"459","author":"T Sato","year":"2009","unstructured":"Sato T, Vries RG, Snippert HJ, van de Wetering M, Barker N, Stange DE, van Es JH, Abo A, Kujala P, Peters PJ, Clevers H (2009) Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche. Nature 459:262\u2013265","journal-title":"Nature"},{"key":"1283_CR17","doi-asserted-by":"publisher","first-page":"1225","DOI":"10.1038\/nm.2470","volume":"17","author":"P Jung","year":"2011","unstructured":"Jung P, Sato T, Merlos-Su\u00e1rez A, Barriga FM, Iglesias M, Rossell D, Auer H, Gallardo M, Blasco MA, Sancho E, Clevers H, Batlle E (2011) Isolation and in vitro expansion of human colonic stem cells. Nat Med 17:1225\u20131227","journal-title":"Nat Med"},{"key":"1283_CR18","doi-asserted-by":"publisher","first-page":"357","DOI":"10.1016\/j.yexcr.2005.02.022","volume":"306","author":"D Pinto","year":"2005","unstructured":"Pinto D, Clevers H (2005) Wnt control of stem cells and differentiation in the intestinal epithelium. Exp Cell Res 306:357\u2013363","journal-title":"Exp Cell Res"},{"key":"1283_CR19","doi-asserted-by":"publisher","first-page":"1190","DOI":"10.1126\/science.1234852","volume":"340","author":"T Sato","year":"2013","unstructured":"Sato T, Clevers H (2013) Growing self-organizing mini-guts from a single intestinal stem cell: mechanism and applications. Science 340:1190\u20131194","journal-title":"Science"},{"key":"1283_CR20","doi-asserted-by":"publisher","first-page":"1138","DOI":"10.1016\/j.stemcr.2022.03.007","volume":"17","author":"YH Tsai","year":"2022","unstructured":"Tsai YH, Wu A, Wu JH, Capeling MM, Holloway EM, Huang S, Czerwinkski M, Glass I, Higgins PDR, Spence JR (2022) Acquisition of NOTCH dependence is a hallmark of human intestinal stem cell maturation. Stem Cell Reports 17:1138\u20131153","journal-title":"Stem Cell Reports"},{"key":"1283_CR21","doi-asserted-by":"publisher","first-page":"959","DOI":"10.1038\/nature03659","volume":"435","author":"JH van Es","year":"2005","unstructured":"van Es JH, van Gijn ME, Riccio O, van den Born M, Vooijs M, Begthel H, Cozijnsen M, Robine S, Winton DJ, Radtke F, Clevers H (2005) Notch\/gamma-secretase inhibition turns proliferative cells in intestinal crypts and adenomas into goblet cells. Nature 435:959\u2013963","journal-title":"Nature"},{"key":"1283_CR22","doi-asserted-by":"publisher","first-page":"49","DOI":"10.1038\/s41421-018-0051-0","volume":"4","author":"Y Li","year":"2018","unstructured":"Li Y, Liu Y, Liu B, Wang J, Wei S, Qi Z, Wang S, Fu W, Chen YG (2018) A growth factor-free culture system underscores the coordination between Wnt and BMP signaling in Lgr5+ intestinal stem cell maintenance. Cell Discov 4:49","journal-title":"Cell Discov"},{"key":"1283_CR23","doi-asserted-by":"publisher","first-page":"1762","DOI":"10.1053\/j.gastro.2011.07.050","volume":"141","author":"T Sato","year":"2011","unstructured":"Sato T, Stange DE, Ferrante M, Vries RG, Van Es JH, Van den Brink S, Van Houdt WJ, Pronk A, Van Gorp J, Siersema PD, Clevers H (2011) Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and Barrett\u2019s epithelium. Gastroenterology 141:1762\u20131772","journal-title":"Gastroenterology"},{"key":"1283_CR24","doi-asserted-by":"publisher","DOI":"10.1016\/j.tiv.2019.104606","volume":"61","author":"E T\u00f6pfer","year":"2019","unstructured":"T\u00f6pfer E, Pasotti A, Telopoulou A, Italiani P, Boraschi D, Ewart M-A, Wilde C (2019) Bovine colon organoids: From 3D bioprinting to cryopreserved multi-well screening platforms. Toxicol In Vitro 61:104606","journal-title":"Toxicol In Vitro"},{"key":"1283_CR25","doi-asserted-by":"publisher","first-page":"911","DOI":"10.1136\/gutjnl-2013-306651","volume":"64","author":"KL VanDussen","year":"2015","unstructured":"VanDussen KL, Marinshaw JM, Shaikh N, Miyoshi H, Moon C, Tarr PI, Ciorba MA, Stappenbeck TS (2015) Development of an enhanced human gastrointestinal epithelial culture system to facilitate patient-based assays. Gut 64:911\u2013920","journal-title":"Gut"},{"key":"1283_CR26","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2020.547102","volume":"11","author":"SS Wilson","year":"2020","unstructured":"Wilson SS, Mayo M, Melim T, Knight H, Patnaude L, Wu X, Phillips L, Westmoreland S, Dunstan R, Fiebiger E, Terrillon S (2020) Optimized culture conditions for improved growth and functional differentiation of mouse and human colon organoids. Front Immunol 11:547102","journal-title":"Front Immunol"},{"key":"1283_CR27","doi-asserted-by":"publisher","DOI":"10.1016\/j.scr.2020.101980","volume":"48","author":"E Mussard","year":"2020","unstructured":"Mussard E, Pouzet C, Helies V, Pascal G, Fourre S, Cherbuy C, Rubio A, Vergnolle N, Combes S, Beaumont M (2020) Culture of rabbit caecum organoids by reconstituting the intestinal stem cell niche in vitro with pharmacological inhibitors or L-WRN conditioned medium. Stem Cell Res 48:101980","journal-title":"Stem Cell Res"},{"key":"1283_CR28","doi-asserted-by":"publisher","first-page":"95","DOI":"10.1038\/s41419-020-03379-9","volume":"12","author":"AM Kip","year":"2021","unstructured":"Kip AM, Soons Z, Mohren R, Duivenvoorden AAM, R\u00f6th AAJ, Cillero-Pastor B, Neumann UP, Dejong CHC, Heeren RMA, Olde Damink SWM, Lenaerts K (2021) Proteomics analysis of human intestinal organoids during hypoxia and reoxygenation as a model to study ischemia-reperfusion injury. Cell Death Dis 12:95","journal-title":"Cell Death Dis"},{"key":"1283_CR29","doi-asserted-by":"publisher","first-page":"215","DOI":"10.1177\/0261192920974026","volume":"48","author":"B Seeger","year":"2020","unstructured":"Seeger B (2020) Farm animal-derived models of the intestinal epithelium: recent advances and future applications of intestinal organoids. Altern Lab Anim 48:215\u2013233","journal-title":"Altern Lab Anim"},{"key":"1283_CR30","doi-asserted-by":"publisher","first-page":"195","DOI":"10.1016\/j.biomaterials.2018.12.006","volume":"194","author":"S Rahmani","year":"2019","unstructured":"Rahmani S, Breyner NM, Su H-M, Verdu EF, Didar TF (2019) Intestinal organoids: a new paradigm for engineering intestinal epithelium in vitro. Biomaterials 194:195\u2013214","journal-title":"Biomaterials"},{"key":"1283_CR31","doi-asserted-by":"publisher","DOI":"10.1002\/cpim.106","volume":"130","author":"C Pleguezuelos-Manzano","year":"2020","unstructured":"Pleguezuelos-Manzano C, Puschhof J, van den Brink S, Geurts V, Beumer J, Clevers H (2020) Establishment and culture of human intestinal organoids derived from adult stem cells. Curr Protoc Immunol 130:e106","journal-title":"Curr Protoc Immunol"},{"key":"1283_CR32","doi-asserted-by":"publisher","first-page":"113","DOI":"10.1016\/j.jcmgh.2017.10.007","volume":"5","author":"Y Wang","year":"2018","unstructured":"Wang Y, Kim R, Gunasekara DB, Reed MI, DiSalvo M, Nguyen DL, Bultman SJ, Sims CE, Magness ST, Allbritton NL (2018) Formation of human colonic crypt array by application of chemical gradients across a shaped epithelial monolayer. Cell Mol Gastroenterol Hepatol 5:113\u2013130","journal-title":"Cell Mol Gastroenterol Hepatol"},{"key":"1283_CR33","doi-asserted-by":"publisher","first-page":"2471","DOI":"10.1038\/nprot.2013.153","volume":"8","author":"H Miyoshi","year":"2013","unstructured":"Miyoshi H, Stappenbeck TS (2013) In vitro expansion and genetic modification of gastrointestinal stem cells in spheroid culture. Nat Protoc 8:2471\u20132482","journal-title":"Nat Protoc"},{"key":"1283_CR34","doi-asserted-by":"publisher","first-page":"822","DOI":"10.3390\/cells9040822","volume":"9","author":"N Kramer","year":"2020","unstructured":"Kramer N, Pratscher B, Meneses AMC, Tschulenk W, Walter I, Swoboda A, Kruitwagen HS, Schneeberger K, Penning LC, Spee B, Kieslinger M, Brandt S, Burgener IA (2020) Generation of differentiating and long-living intestinal organoids reflecting the cellular diversity of canine intestine. Cells 9:822","journal-title":"Cells"},{"key":"1283_CR35","doi-asserted-by":"publisher","first-page":"C794","DOI":"10.1152\/ajpcell.00420.2020","volume":"320","author":"AM Barnett","year":"2021","unstructured":"Barnett AM, Mullaney JA, Hendriks C, Le Borgne L, McNabb WC, Roy NC (2021) Porcine colonoids and enteroids keep the memory of their origin during regeneration. Am J Physiol Cell Physiol 320:C794\u2013C805","journal-title":"Am J Physiol Cell Physiol"},{"key":"1283_CR36","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0066465","volume":"8","author":"LM Gonzalez","year":"2013","unstructured":"Gonzalez LM, Williamson I, Piedrahita JA, Blikslager AT, Magness ST (2013) Cell lineage identification and stem cell culture in a porcine model for the study of intestinal epithelial regeneration. PLoS ONE 8:e66465","journal-title":"PLoS ONE"},{"key":"1283_CR37","doi-asserted-by":"publisher","first-page":"54","DOI":"10.1186\/s13567-018-0547-5","volume":"49","author":"CA Hamilton","year":"2018","unstructured":"Hamilton CA, Young R, Jayaraman S, Sehgal A, Paxton E, Thomson S, Katzer F, Hope J, Innes E, Morrison LJ, Mabbott NA (2018) Development of in vitro enteroids derived from bovine small intestinal crypts. Vet Res 49:54","journal-title":"Vet Res"},{"key":"1283_CR38","doi-asserted-by":"publisher","first-page":"123","DOI":"10.1007\/s00441-016-2367-0","volume":"365","author":"HA Khalil","year":"2016","unstructured":"Khalil HA, Lei NY, Brinkley G, Scott A, Wang J, Kar UK, Jabaji ZB, Lewis M, Mart\u00edn MG, Dunn JC, Stelzner MG (2016) A novel culture system for adult porcine intestinal crypts. Cell Tissue Res 365:123\u2013134","journal-title":"Cell Tissue Res"},{"key":"1283_CR39","doi-asserted-by":"publisher","first-page":"377","DOI":"10.1038\/s42003-021-01901-z","volume":"4","author":"TJ Nash","year":"2021","unstructured":"Nash TJ, Morris KM, Mabbott NA, Vervelde L (2021) Inside-out chicken enteroids with leukocyte component as a model to study host-pathogen interactions. Commun Biol 4:377","journal-title":"Commun Biol"},{"key":"1283_CR40","doi-asserted-by":"publisher","DOI":"10.3389\/fvets.2022.921160","volume":"9","author":"R Blake","year":"2022","unstructured":"Blake R, Jensen K, Mabbott N, Hope J, Stevens J (2022) The development of 3D bovine intestinal organoid derived models to investigate Mycobacterium Avium ssp Paratuberculosis pathogenesis. Front Vet Sci 9:921160","journal-title":"Front Vet Sci"},{"key":"1283_CR41","doi-asserted-by":"publisher","first-page":"15","DOI":"10.1186\/s13567-022-01033-0","volume":"53","author":"KM Sutton","year":"2022","unstructured":"Sutton KM, Orr B, Hope J, Jensen SR, Vervelde L (2022) Establishment of bovine 3D enteroid-derived 2D monolayers. Vet Res 53:15","journal-title":"Vet Res"},{"key":"1283_CR42","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0256143","volume":"16","author":"P Hoffmann","year":"2021","unstructured":"Hoffmann P, Schnepel N, Langeheine M, K\u00fcnnemann K, Grassl GA, Brehm R, Seeger B, Mazzuoli-Weber G, Breves G (2021) Intestinal organoid-based 2D monolayers mimic physiological and pathophysiological properties of the pig intestine. PLoS ONE 16:e0256143","journal-title":"PLoS ONE"},{"key":"1283_CR43","doi-asserted-by":"publisher","first-page":"375","DOI":"10.3389\/fcell.2020.00375","volume":"8","author":"B van der Hee","year":"2020","unstructured":"van der Hee B, Madsen O, Vervoort J, Smidt H, Wells JM (2020) Congruence of transcription programs in adult stem cell-derived jejunum organoids and original tissue during long-term culture. Front Cell Dev Biol 8:375","journal-title":"Front Cell Dev Biol"},{"key":"1283_CR44","doi-asserted-by":"publisher","first-page":"241","DOI":"10.1111\/evj.12734","volume":"50","author":"A Stieler Stewart","year":"2018","unstructured":"Stieler Stewart A, Freund JM, Gonzalez LM (2018) Advanced three-dimensional culture of equine intestinal epithelial stem cells. Equine Vet J 50:241\u2013248","journal-title":"Equine Vet J"},{"key":"1283_CR45","doi-asserted-by":"publisher","first-page":"108","DOI":"10.1186\/s13567-021-00976-0","volume":"52","author":"S Hellman","year":"2021","unstructured":"Hellman S (2021) Generation of equine enteroids and enteroid-derived 2D monolayers that are responsive to microbial mimics. Vet Res 52:108","journal-title":"Vet Res"},{"key":"1283_CR46","doi-asserted-by":"publisher","first-page":"445","DOI":"10.1186\/s12917-022-03552-6","volume":"18","author":"AS Stewart","year":"2022","unstructured":"Stewart AS, Schaaf CR, Veerasammy B, Freund JM, Gonzalez LM (2022) Culture of equine intestinal epithelial stem cells after delayed tissue storage for future applications. BMC Vet Res 18:445","journal-title":"BMC Vet Res"},{"key":"1283_CR47","doi-asserted-by":"publisher","first-page":"34","DOI":"10.1111\/evj.41_13495","volume":"53","author":"S Hellman","year":"2021","unstructured":"Hellman S (2021) New dimensions of equine intestinal organoids to model host-microbe interactions in vitro. Equine Vet J 53:34","journal-title":"Equine Vet J"},{"key":"1283_CR48","volume":"6","author":"K O\u2019Rourke","year":"2016","unstructured":"O\u2019Rourke K, Ackerman S, Dow L, Lowe S (2016) Isolation, culture, and maintenance of mouse intestinal stem cells. Bio Protoc 6:e1733","journal-title":"Bio Protoc"},{"key":"1283_CR49","doi-asserted-by":"publisher","first-page":"3710836","DOI":"10.1155\/2016\/3710836","volume":"2016","author":"A Pastu\u0142a","year":"2016","unstructured":"Pastu\u0142a A, Middelhoff M, Brandtner A, Tobiasch M, H\u00f6hl B, Nuber AH, Demir IE, Neupert S, Kollmann P, Mazzuoli-Weber G, Quante M (2016) Three-dimensional gastrointestinal organoid culture in combination with nerves or fibroblasts: a method to characterize the gastrointestinal stem cell niche. Stem Cells Int 2016:3710836","journal-title":"Stem Cells Int"},{"key":"1283_CR50","doi-asserted-by":"publisher","first-page":"6356","DOI":"10.18632\/oncotarget.23721","volume":"9","author":"MO Nam","year":"2018","unstructured":"Nam MO, Hahn S, Jee JH, Hwang TS, Yoon H, Lee DH, Kwon MS, Yoo J (2018) Effects of a small molecule R-spondin-1 substitute RS-246204 on a mouse intestinal organoid culture. Oncotarget 9:6356\u20136368","journal-title":"Oncotarget"},{"key":"1283_CR51","doi-asserted-by":"publisher","first-page":"177","DOI":"10.1016\/j.stem.2016.11.001","volume":"20","author":"O Basak","year":"2017","unstructured":"Basak O, Beumer J, Wiebrands K, Seno H, van Oudenaarden A, Clevers H (2017) Induced quiescence of Lgr5+ stem cells in intestinal organoids enables differentiation of hormone-producing enteroendocrine cells. Cell Stem Cell 20:177-190.e4","journal-title":"Cell Stem Cell"},{"key":"1283_CR52","doi-asserted-by":"publisher","first-page":"676","DOI":"10.1038\/nmeth.2019","volume":"9","author":"J Schindelin","year":"2012","unstructured":"Schindelin J, Arganda-Carreras I, Frise E, Kaynig V, Longair M, Pietzsch T, Preibisch S, Rueden C, Saalfeld S, Schmid B, Tinevez JY, White DJ, Hartenstein V, Eliceiri K, Tomancak P, Cardona A (2012) Fiji: an open-source platform for biological-image analysis. Nat Methods 9:676\u2013682","journal-title":"Nat Methods"},{"key":"1283_CR53","doi-asserted-by":"publisher","first-page":"947482","DOI":"10.3389\/fvets.2022.947482","volume":"9","author":"F Dengler","year":"2022","unstructured":"Dengler F, Sternberg F, Grages M, K\u00e4stner SB, Verhaar N (2022) Adaptive mechanisms in no flow vs low flow ischemia in equine Jejunum epithelium: different paths to the same destination. Front Vet Sci 9:947482","journal-title":"Front Vet Sci"},{"key":"1283_CR54","doi-asserted-by":"publisher","first-page":"611","DOI":"10.1373\/clinchem.2008.112797","volume":"55","author":"SA Bustin","year":"2009","unstructured":"Bustin SA, Benes V, Garson JA, Hellemans J, Huggett J, Kubista M, Mueller R, Nolan T, Pfaffl MW, Shipley GL, Vandesompele J, Wittwer CT (2009) The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem 55:611\u2013622","journal-title":"Clin Chem"},{"key":"1283_CR55","doi-asserted-by":"publisher","first-page":"125","DOI":"10.1007\/s10142-023-01055-7","volume":"23","author":"F Xie","year":"2023","unstructured":"Xie F, Wang J, Zhang B (2023) RefFinder: a web-based tool for comprehensively analyzing and identifying reference genes. Funct Integr Genomics 23:125","journal-title":"Funct Integr Genomics"},{"key":"1283_CR56","doi-asserted-by":"publisher","first-page":"15","DOI":"10.1053\/j.gastro.2009.05.035","volume":"137","author":"LG van der Flier","year":"2009","unstructured":"van der Flier LG, Haegebarth A, Stange DE, van de Wetering M, Clevers H (2009) OLFM4 is a robust marker for stem cells in human intestine and marks a subset of colorectal cancer cells. Gastroenterology 137:15\u201317","journal-title":"Gastroenterology"},{"key":"1283_CR57","first-page":"698","volume":"6","author":"RH Powell","year":"2017","unstructured":"Powell RH, Behnke MS (2017) WRN conditioned media is sufficient for in vitro propagation of intestinal organoids from large farm and small companion animals. Biol Open 6:698\u2013705","journal-title":"Biol Open"},{"key":"1283_CR58","doi-asserted-by":"publisher","first-page":"153","DOI":"10.1016\/j.bbrc.2022.11.008","volume":"637","author":"S Ohira","year":"2022","unstructured":"Ohira S, Yokoi Y, Ayabe T, Nakamura K (2022) Efficient and simple genetic engineering of enteroids using mouse isolated crypts for investigating intestinal functions. Biochem Biophys Res Commun 637:153\u2013160","journal-title":"Biochem Biophys Res Commun"},{"key":"1283_CR59","doi-asserted-by":"publisher","first-page":"289","DOI":"10.1146\/annurev-physiol-030212-183744","volume":"75","author":"HC Clevers","year":"2013","unstructured":"Clevers HC, Bevins CL (2013) Paneth cells: maestros of the small intestinal crypts. Annu Rev Physiol 75:289\u2013311","journal-title":"Annu Rev Physiol"},{"key":"1283_CR60","doi-asserted-by":"publisher","first-page":"225","DOI":"10.3390\/v10050225","volume":"10","author":"M Holly","year":"2018","unstructured":"Holly M, Smith J (2018) Paneth cells during viral infection and pathogenesis. Viruses 10:225","journal-title":"Viruses"},{"key":"1283_CR61","doi-asserted-by":"publisher","first-page":"393","DOI":"10.3390\/nu15020393","volume":"15","author":"IH de Lange","year":"2023","unstructured":"de Lange IH, van Gorp C, Massy KRI, Kessels L, Kloosterboer N, Bj\u00f8rnshave A, Stampe Ostenfeld M, Damoiseaux JGMC, Derikx JPM, van Gemert WG, Wolfs TGAM (2023) Hypoxia-driven changes in a human intestinal organoid model and the protective effects of hydrolyzed whey. Nutrients 15:393","journal-title":"Nutrients"},{"key":"1283_CR62","doi-asserted-by":"publisher","first-page":"19","DOI":"10.1186\/s12915-018-0481-z","volume":"16","author":"SC Pearce","year":"2018","unstructured":"Pearce SC, Al-Jawadi A, Kishida K, Yu S, Hu M, Fritzky LF, Edelblum KL, Gao N, Ferraris RP (2018) Marked differences in tight junction composition and macromolecular permeability among different intestinal cell types. BMC Biol 16:19","journal-title":"BMC Biol"},{"key":"1283_CR63","doi-asserted-by":"publisher","first-page":"33","DOI":"10.1186\/s12915-019-0652-6","volume":"17","author":"L Chandra","year":"2019","unstructured":"Chandra L, Borcherding DC, Kingsbury D, Atherly T, Ambrosini YM, Bourgois-Mochel A, Yuan W, Kimber M, Qi Y, Wang Q, Wannemuehler M, Ellinwood NM, Snella E, Martin M, Skala M, Meyerholz D, Estes M, Fernandez-Zapico ME, Jergens AE, Mochel JP, Allenspach K (2019) Derivation of adult canine intestinal organoids for translational research in gastroenterology. BMC Biol 17:33","journal-title":"BMC Biol"},{"key":"1283_CR64","doi-asserted-by":"publisher","first-page":"1003","DOI":"10.1038\/nature06196","volume":"449","author":"N Barker","year":"2007","unstructured":"Barker N, van Es JH, Kuipers J, Kujala P, van den Born M, Cozijnsen M, Haegebarth A, Korving J, Begthel H, Peters PJ, Clevers H (2007) Identification of stem cells in small intestine and colon by marker gene Lgr5. Nature 449:1003\u20131007","journal-title":"Nature"},{"key":"1283_CR65","doi-asserted-by":"publisher","first-page":"358","DOI":"10.2460\/ajvr.76.4.358","volume":"76","author":"LM Gonzalez","year":"2015","unstructured":"Gonzalez LM, Kinnin LA, Blikslager AT (2015) Characterization of discrete equine intestinal epithelial cell lineages. Am J Vet Res 76:358\u2013366","journal-title":"Am J Vet Res"},{"key":"1283_CR66","doi-asserted-by":"publisher","first-page":"261","DOI":"10.1038\/s41467-021-27901-5","volume":"13","author":"D Zeve","year":"2022","unstructured":"Zeve D, Stas E, de Sousa CJ, Mannam P, Qi W, Yin X, Dubois S, Shah MS, Syverson EP, Hafner S, Karp JM, Carlone DL, Ordovas-Montanes J, Breault DT (2022) Robust differentiation of human enteroendocrine cells from intestinal stem cells. Nat Commun 13:261","journal-title":"Nat Commun"},{"key":"1283_CR67","doi-asserted-by":"publisher","first-page":"356","DOI":"10.1016\/j.devcel.2020.12.020","volume":"56","author":"LE Sanman","year":"2021","unstructured":"Sanman LE, Chen IW, Bieber JM, Steri V, Trentesaux C, Hann B, Klein OD, Wu LF, Altschuler SJ (2021) Transit-amplifying cells coordinate changes in intestinal epithelial cell-type composition. Dev Cell 56:356-365.e9","journal-title":"Dev Cell"},{"key":"1283_CR68","doi-asserted-by":"publisher","first-page":"610","DOI":"10.1136\/gutjnl-2012-304067","volume":"63","author":"A Reynolds","year":"2014","unstructured":"Reynolds A, Wharton N, Parris A, Mitchell E, Sobolewski A, Kam C, Bigwood L, El Hadi A, M\u00fcnsterberg A, Lewis M, Speakman C, Stebbings W, Wharton R, Sargen K, Tighe R, Jamieson C, Hernon J, Kapur S, Oue N, Yasui W, Williams MR (2014) Canonical Wnt signals combined with suppressed TGF\u03b2\/BMP pathways promote renewal of the native human colonic epithelium. Gut 63:610\u2013621","journal-title":"Gut"},{"key":"1283_CR69","doi-asserted-by":"publisher","first-page":"14700","DOI":"10.1073\/pnas.0702305104","volume":"104","author":"ME Binnerts","year":"2007","unstructured":"Binnerts ME, Kim KA, Bright JM, Patel SM, Tran K, Zhou M, Leung JM, Liu Y, Lomas WE 3rd, Dixon M, Hazell SA, Wagle M, Nie WS, Tomasevic N, Williams J, Zhan X, Levy MD, Funk WD, Abo A (2007) R-Spondin1 regulates Wnt signaling by inhibiting internalization of LRP6. Proc Natl Acad Sci USA 104:14700\u201314705","journal-title":"Proc Natl Acad Sci USA"},{"key":"1283_CR70","doi-asserted-by":"publisher","first-page":"415","DOI":"10.1038\/nature09637","volume":"469","author":"T Sato","year":"2011","unstructured":"Sato T, van Es JH, Snippert HJ, Stange DE, Vries RG, van den Born M, Barker N, Shroyer NF, van de Wetering M, Clevers H (2011) Paneth cells constitute the niche for Lgr5 stem cells in intestinal crypts. Nature 469:415\u2013418","journal-title":"Nature"},{"key":"1283_CR71","doi-asserted-by":"publisher","DOI":"10.15252\/embr.202256454","volume":"24","author":"L Berkov\u00e1","year":"2023","unstructured":"Berkov\u00e1 L, Fazilaty H, Yang Q, Kubov\u010diak J, Stastna M, Hrckulak D, Vojtechova M, Dalessi T, Br\u00fcgger MD, Hausmann G, Liberali P, Korinek V, Basler K, Valenta T (2023) Terminal differentiation of villus tip enterocytes is governed by distinct Tgf\u03b2 superfamily members. EMBO Rep 24:e56454","journal-title":"EMBO Rep"},{"key":"1283_CR72","doi-asserted-by":"publisher","first-page":"887","DOI":"10.1053\/j.gastro.2007.06.066","volume":"133","author":"BA Auclair","year":"2007","unstructured":"Auclair BA, Benoit YD, Rivard N, Mishina Y, Perreault N (2007) Bone morphogenetic protein signaling is essential for terminal differentiation of the intestinal secretory cell lineage. Gastroenterology 133:887\u2013896","journal-title":"Gastroenterology"},{"key":"1283_CR73","doi-asserted-by":"publisher","first-page":"3535","DOI":"10.1038\/s41598-020-60521-5","volume":"10","author":"W Dieterich","year":"2020","unstructured":"Dieterich W, Neurath MF, Zopf Y (2020) Intestinal ex vivo organoid culture reveals altered programmed crypt stem cells in patients with celiac disease. Sci Rep 10:3535","journal-title":"Sci Rep"},{"key":"1283_CR74","volume":"181","author":"V Gabriel","year":"2022","unstructured":"Gabriel V, Zdyrski C, Sahoo DK, Dao K, Bourgois-Mochel A, Atherly T, Martinez MN, Volpe DA, Kopper J, Allenspach K, Mochel JP (2022) Canine intestinal organoids in a dual-chamber permeable support system. J Vis Exp 181:e63612","journal-title":"J Vis Exp"}],"container-title":["Veterinary Research"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s13567-024-01283-0.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1186\/s13567-024-01283-0\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s13567-024-01283-0.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,3,16]],"date-time":"2024-03-16T06:05:06Z","timestamp":1710569106000},"score":1,"resource":{"primary":{"URL":"https:\/\/veterinaryresearch.biomedcentral.com\/articles\/10.1186\/s13567-024-01283-0"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,3,16]]},"references-count":74,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2024,12]]}},"alternative-id":["1283"],"URL":"https:\/\/doi.org\/10.1186\/s13567-024-01283-0","relation":{},"ISSN":["1297-9716"],"issn-type":[{"value":"1297-9716","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,3,16]]},"assertion":[{"value":"21 November 2023","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"8 February 2024","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"16 March 2024","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The authors declare that they have no competing interests.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"30"}}